Anti-Tau (phospho S199) antibody [EPR2401Y]

• WB

Lab

Western blot - Anti-Tau (phospho S199) antibody [EPR2401Y] (AB81268)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Tau (phospho S199) antibody [EPR2401Y] (ab81268) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a flag tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human Tau expression vector containing a flag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human MAP2 expression vector containing a flag whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human MAP4 expression vector containing a flag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 78 kDa

Observed band size: 55-100 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-Tau (phospho S199) antibody [EPR2401Y] (AB81268)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Tau (phospho S199) antibody [EPR2401Y] (ab81268) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 10 µg

Lane 2:

Human hippocampus tissue lysate. The membrane was incubated with phosphatase at 10 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/2000 dilution

Predicted band size: 78 kDa

Observed band size: 55 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-Tau (phospho S199) antibody [EPR2401Y] (AB81268)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Tau (phospho S199) antibody [EPR2401Y] (ab81268) at 1/1000 dilution

Lane 1:

Mouse cerebral cortex tissue lysate at 10 µg

Lane 2:

Mouse cerebral cortex tissue lysate, The membrane was incubated with phosphatase at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 78 kDa

Observed band size: 55 kDa

false

Exposure time: 5s

• WB

Unknown

Western blot - Anti-Tau (phospho S199) antibody [EPR2401Y] (AB81268)

All lanes:

Western blot - Anti-Tau (phospho S199) antibody [EPR2401Y] (ab81268) at 1/20000 dilution

Lane 1:

SH SY5Y cell lysate at 10 µg

Lane 2:

SH SY5Y cell lysate treated with alkaline phosphatase at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 78 kDa

Observed band size: 55 kDa

false

• Dot

Unknown

Dot Blot - Anti-Tau (phospho S199) antibody [EPR2401Y] (AB81268)

Dot blot analysis of Tau (pS199) peptide (Lane 1) and Tau non-phospho peptide (Lane 2) labelling Tau (pS199) with ab81268 at a dilution of 1/1000. ab97051 (peroxidase-conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

Exposure time : 3 minutes.
Blocking and dilution buffer : 5% NFDM/TBST.

• Dot

Supplier Data

Dot Blot - Anti-Tau (phospho S199) antibody [EPR2401Y] (AB81268)

Dot blot analysis using 1/1000 dilution ab81268 and Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary at 1/100000 dilution.

Blocking and diluting buffer : 5% NFDM/TBST

Lane 1 : Tau non-phospho peptide

Lane 2 : Tau S199 phospho peptide

Lane 3 : Tau S202 phospho peptide

Lane 4 : Tau S199+S202 phospho peptide

Exposure time : 3 minutes

• WB

CiteAb

Western blot - Anti-Tau (phospho S199) antibody [EPR2401Y] (AB81268)

Tau (phospho S199) western blot using anti-Tau (phospho S199) antibody [EPR2401Y] ab81268. Publication image and figure legend from Yang, S. H., Lee, D. K., et al., 2017, EMBO Mol Med, PubMed 27861127.

ab81268 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab81268 please see the product overview.

Nec‐1 inhibits the phosphorylation and aggregation of tauAWestern blot analysis of phosphorylation levels of tau and expression levels of indicated proteins in the cortical and hippocampal regions of the brain in APP/PS1 mice after administration of Nec‐1 (6.25 mg/kg) or vehicle (2.5% DMSO in PBS). The membranes analysed are identical to those in Fig 5F.BImmunohistochemical analysis of cortical and hippocampal regions of the brains in wild‐type (Wt) and APP/PS1 mice after administration of Nec‐1 (6.25 mg/kg) or vehicle (2.5% DMSO in PBS) for phosphorylated tau expression levels. Anti‐tau (phospho‐Ser199) antibody (red colour) was applied for staining tau phosphorylation on serine‐199 and Hoechst 33342 (blue colour) for nuclear counterstaining in the brain sections. Scale bars = 200 μm. Yellow and red boxes indicate the cortical and hippocampal region of brains, respectively.CWestern blot analyses of phosphorylated RIPK3 expression in cortex and hippocampus of APP/PS1 mice injected with Nec‐1 (upper), and primary cultured astrocytes treated with Nec‐1 (lower). The membranes analysed are identical to those in Figs 1G and 5F.D, EThT assays for the inhibition of tau aggregation (D) and for disaggregation of tau (E). Fluorescence intensity was normalized to tau aggregates (100%, day 5). All data are representative results of at least three independent experiments.FSurface plasmon resonance sensorgrams of Nec‐1 targeting tau (left), and the corresponding dissociation fitting curve from the saturated region (right) under various concentrations of Nec‐1.Data information : In (D and E), data are presented as mean ± SEM. ***P ≤ 0.001 (one‐way ANOVA followed by Bonferroni's post hoc comparisons tests). Exact P‐values are shown in Table EV5. Source data are available online for this figure.

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